In this paper, natural deep eutectic solvents (NADES)-based for liquid phase microextraction (LPME) is proposed for the isolation of different psychoactive substances in water and biological fluids followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The NADES composition was optimized (L-menthol:octanoic acid, 1:2 M ratio) prior to the study of the extraction conditions such as sample and elution volumes, obtaining up to 16-fold preconcentration factor. Next, the quality parameters were studied including linearity from LOQs (0.0006–0.05 μg L−1) to 100 μg L−1, precision values (expressed as relative standard deviation, RSD≤8 %) and recoveries above 70 % in most cases. Certified saliva and serum samples were analyzed by the proposed method to assess the accuracy of the developed procedure, providing values statistically comparable to the certified ones. In addition, the proposed methodology was evaluated by applying green metrics (AGREEprep tool) and the obtained values were compared with those reported for a similar procedure using LPME with common organic solvents such as chloroform or dichloromethane, outperforming them in both cases, which points out the potential of this method from the sustainability point of view.